Advertisement

Clofibrate Selectively Induces Azoreduction of Dimethylaminoazobenzene (DAB) by Rat Liver Microsomes

  • W. G. Levine
  • H. Raza
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 197)

Abstract

Reduction of azo linkages in xenobiotics has long been known in biological systems. Fifty years ago, the antibacterial dye, prontosil, was shown to be reduced to the active product, sulfanilamide, by colonic bacterial. Hernandez et a1.2 found that reduction of prontosil and neotetrazolium by rat liver microsomes is partially inhibited by CO, implying the involvement of cytochrome P-450. Purified NADPH-cytochrome c reductase also catalyzes reduction and accounts for CO-insensitive activity3. Fujita and Peisach4 reported that azoreduction of amaranth is almost completely inhibited by CO and is induced by treatment with phenobarbital (PB) or 3-methylcholanthrene (MC). Under all conditions, the rate of azoreduction of amaranth is proportional to total microsomal cytochrome P-450. Antibodies against PB- and MC-induced cytochrome P-450 almost completely inhibit azoreduction of amaranth in PB- or MC-induced microsomes, respectively6. Recently, it was shown that purified cytochrome P-450 from PB-induced rat livers readily reduces amaranth7. Thus, there are several distinct azoreductases in microsomes which can be distinguished by their substrate spectificities. FMN and FAD markedly stimulate azoreduction of amaranth5 and neoprontosil8. The stimulated activity is unaffected by CO, implying, although not necessarily proving, that cytochrome P-450 is not involved. However, it was later demonstrated that these flavines could transfer electrons from both NADPH-cytochrome c reductase and cytochrome P-450 to dye.9

Keywords

Lauric Acid Microsomal Cytochrome Reductive Metabolism Hypolipidemic Drug Azoreductase Activity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    J. Trefouel, J. Trefouel, F. Netti, and D. Bovet, Activite de paminophenylsulfamide sur les infections streptococciques experimentales de la souris et du lapin, C.R. Seanc. Soc. Biol., 120: 756 (1935).Google Scholar
  2. 2.
    P.H. Hernandez, P. Mazel and J.R. Gillette, Studies on the mechanism of action of mammalian hepatic azoreductase-II: The effects of phenobarbital and 3-methylcholanthrene on carbon monoxide sensitive and insensitive azoreductase activities, Biochem. Pharmacol., 16: 1877 (1967).PubMedCrossRefGoogle Scholar
  3. 3.
    P.H. Hernandez, J.R. Gillette and P. Mazel, Studies on the mechanism of action of mammalian hepatic azoreductase-I: Azoreductase activity of reduced nicotinamide adenine dinucleotidephosphatecytochrome c reductase, Biochem. Pharmacol., 16: 1859 (1967).PubMedCrossRefGoogle Scholar
  4. 4.
    S. Fujita and J. Peisach, Liver microsomal cytochromes P-450 and azoreductase activity, J. Biol. Chem., 252: 4512 (1978).Google Scholar
  5. 5.
    S. Fujita and J. Peisach, The stimulation of microsomal azoreduction by flavins, Biochim. Biophys. Acta. 719: 178 (1982).PubMedCrossRefGoogle Scholar
  6. 6.
    S. Fujita, Y. Okada, and J. Peisach, Inhibition of azoreductase activity by antibodies against cytochrome P-450 and P-448, Biochem Biophys. Res. Cununun., 102: 492 (1391).CrossRefGoogle Scholar
  7. 7.
    A.K. Mallett, L.J. King, and R. Walker, Solubilization purification and reconstitution of hepatic microsomal azoreductase activity, Biochem. Pharmacol., 34: 337 (1985).PubMedCrossRefGoogle Scholar
  8. 8.
    L. Shargel and P. Mazel, Influence of 2,4 dichloro-6-phenoxymethylamine (DPEA) and (3-diethylaminoethyl diphenylpropylacetate (SKF 525-A) on hepatic microsomal azoreductase activity from phenobarbital or 3-methylcholanthrene induced rats, Biochem. Pharmacol., 21: 69 (1972).PubMedCrossRefGoogle Scholar
  9. 9.
    A.K. Mallett, L.J. King, and R. Walker, A continuous spectrophotometric determination of hepatic microsomal azoreductase activity and its dependence on cytochrome P-450, Biochem. J., 201: 589 (1982).PubMedGoogle Scholar
  10. 10.
    M-T. Huang, G.T. Miwa, N. Cronheim, and A.Y.H. Lu, Rat liver cytosolic azoreductase: Electron transport properties and the mechanism of dicumarol inhibition of the purified enzyme, J. Biol Chem., 254: 11223 (1979).PubMedGoogle Scholar
  11. 11.
    T. Zimmerman, H.G. Kulla, and T. Leisinger, Properties of purified Orange II azoreductase, the enzyme initiating azo dye degradation by Pseudomonas KF46, Eur. J. Biochem., 129: 197 (1982).CrossRefGoogle Scholar
  12. 12.
    G.C. Mueller and J.A. Miller, The reductive cleavage of 4-dimethylaminoazobenzene by rat liver: The intracellular distribution of the enzyme system and its requirement for triphosphopyridine nucleotide, J. Biol. Chem., 180: 1125 (1949).PubMedGoogle Scholar
  13. 13.
    A.H. Conney, E.C. Miller, and J.A. Miller, The metabolism of methylated aminoazo dyes. V. Evidence for induction of enzyme synthesis in the rat by 3-methylcholanthrene, Cancer Res., 16: 450 (1956).PubMedGoogle Scholar
  14. 14.
    R.P. Mason, F.J. Peterson, and J.L. Holtzman, Inhibition of azoreductase by oxygen: the role of the azo anion free radical metabolite in the reduction of oxygen to superoxide, Mol. Pharmacol., 14: 665 (1978).PubMedGoogle Scholar
  15. 15.
    H. Autrup and G.P. Warwick, Some characteristics of two azoreductase systems in rat liver. Relevance to the activity of 2-[4’-di(2’bromopropyl)-aminophenylazo] benzoic acid (CB10–252), a compound possessing latent cytotoxic activity, Chem. Biol. Interact., 11: 329 (1975).PubMedCrossRefGoogle Scholar
  16. 16.
    F. De Cloitre, J. Chauveau, and M. Martin, Influence of age and 3methylcholanthrene on azodye carcinogenesis and metabolism of pdimethylaminoazobenzene in rat liver, Int. J. Cancer, 11: 676 (1973).CrossRefGoogle Scholar
  17. 17.
    P.S. De-Araujo, E. De-Andrade-Silva, and I. Raw, Effect of drugs and hormones on rat liver dimethylaminoazobenzene reductase activity, Braz. J. Med. Biol. Res., 15: 17 (1982).PubMedGoogle Scholar
  18. 18.
    B.M. Elliot, Azoreductase activity of Sprague Dawley and Wistarderived rats towards both carcinogenic and non-carcinogenic analogues of 4-dimethylaminophenylazobenzene (DAB). Carcinogenesis, 5: 1051 (1984).CrossRefGoogle Scholar
  19. 19.
    B. Ketterer, P. Ross-Mansell, and H. Davidson, The effect of the administration of N,N-dimethyl-4-amino-azobenzene (DAB) on the activity of DAB-reductase and NADPH-cytochrome c reductase, Chem. Biol. Interact., 2: 183 (1970).PubMedCrossRefGoogle Scholar
  20. 20.
    W.G. Levine, Studies on microsomal azoreduction of N,N-dimethyl-4aminoazobenzene (DAB) and its derivatives, Biochem. Pharmacol.,in press (1985).Google Scholar
  21. 21.
    W.G. Levine and S.B. Lee, Effect of glutathione on the metabolism of N,N-dimethyl-4-aminoazobenzene by rat liver microsomes, Drug Metab. Dispos., 11: 239 (1983).PubMedGoogle Scholar
  22. 22.
    J.K. Reddy and N.D. Lalwani, Carcinogenesis by hepatic peroxisome proliferators: Evaluation of the risk of hypolipidemic drugs and industrial plasticizers to humans, CRC Crit. Rev. Toxicol., 12: 1 (1983).CrossRefGoogle Scholar
  23. 23.
    T.C. Orton and G.L. Parker, The effect of hypolipidemic agents on the hepatic microsomal drug metabolizing enzyme system of the rat. Induction of cytochrome (s) P-450 with specificity toward terminal hydroxylation of lauric acid, Drug Metab. Dispos., 10: 110 (1982).PubMedGoogle Scholar
  24. 24.
    P.P. Tamburini, H.A. Masson, S.K. Bains, R.J. Makowski, B. Morris and G.G. Gibson, Multiple forms of hepatic cytochrome P-450. Purification, characterization and comparison of a novel clofibrateinduced isazyme with other major forms of cytochrome P-450, Eur. J. Biochem. 139: 235 (1984).PubMedCrossRefGoogle Scholar
  25. 25.
    G.L. Parker, and T.C.Orton, Induction by oxyisobutyrates of hepatic and kidney microsomal cytochrome P-450 with specificity towards hydroxylation of fatty acids, in “Biochemistry, Biophysics and Regulation of Cytochrome P-450,” J.A. Gustafsson, J.C. Duke, A. Mode and J. Rafter, eds. Elsevier/North Holland Press. Amsterdam (1980).Google Scholar
  26. 26.
    R.A. Salvador, S. Haber, C. Atkins, B.W. Gommi and R.M. Welch, Effect of clofibrate and 1-methyl-4-piperidylbis (p-chlorophenoxy) acetate (Sandoz 42–348) on steroid and drug metabolism by rat liver microsomes, Life Sci. 9 part 1I: 397 (1970).CrossRefGoogle Scholar
  27. 27.
    W.G. Levine and A.Y.H. Luu, Rleof isozymes of cytochrome P-450 in the metabolism of N,N-dimethyl-4-aminoazobenzene in the rat, Drug Metab. Dispos. 10: 102 (1982).PubMedGoogle Scholar
  28. 28.
    H. Lowry, N.J. Rosebrough, A.L. Farr, and R.J. Randall, Protein measurement with the Folin phenol reagent, J. Biol. Chem. 193: 265 (1951).PubMedGoogle Scholar
  29. 29.
    J.D. Dignam and H.W. Strobel, NADPH-cytochrome P-450 reductase from rat liver: Purification by affinity chromatography and characterization, Biochemistry 16: 1116 (1977).PubMedCrossRefGoogle Scholar
  30. 30.
    F.J. Weibel, J.C. Lentz, L. Diamond, and H.V. Gelboin, Aryl hydrocarbon (benzo[a]pyrene) hydroxylase in microsomes from rat tissues: differential inhibition and stimulation by benzoflavones and organic solvents, Arch. Biochem. Biophys., 144: 78 (1971).CrossRefGoogle Scholar
  31. 31.
    F. Mitani, E.A. Shepard, I.R. Phillips, and B.R. Rabin, Complexes of cytochrome P-450 with metyrapone. A convenient method for the quantitative analysis of phenobarbital-inducible cytochrome P450 in rat liver microsomes, FEBS Lett 148: 302 (1982).PubMedCrossRefGoogle Scholar
  32. 32.
    R.T. Okita and B.S.S. Masters, Effect of phenobarbital treated and cytochrome P-450 inhibitors on the laurate omega and (omega -1)hydroxylase activities of rat liver microsomes, Drug Metab. Dispos. 8: 147 (1980).PubMedGoogle Scholar
  33. 33.
    R. Walker, The metabolism of azo compounds: A review of the literature, Fd. Cosmet. Toxicol., 8: 659 (1970).CrossRefGoogle Scholar
  34. 34.
    K-T. Chung, The significance of azo-reduction in the mutagenesis and carcinogenesis of azo dyes, Mut. Res. 14: 269 (1983).Google Scholar
  35. 35.
    A.Y.H. Lu and S.B. West, Multiplicity of mammalian microsomal cytochromes P-450, Pharmacol. Rev., 31: 277 (1980).Google Scholar
  36. 36.
    E.G. Schuetz, S.A. Wrighton, J.L. Barwick, and P.S. Guzelian, Induction of cytochrome P-450 by glucocorticoids in rat liver. I. Evidence that glucocorticoids and pregnenalone 16a-carbonitrile regulate de novo synthesis of a common form of cytochrome P-450 in cultures of adult rat hepatocytes and in the liver in vivo, J. Biol. Chem., 259: 1999 (1984).PubMedGoogle Scholar
  37. 37.
    T.R. Fennell, M. Dickins, and J.W. Bridges, Interaction of isosafrole in vivo with rat hepatic microsomal cytochrome P-450 following treatment with phenobarbitone or 20-methylcholanthrene, Biochem. Pharmacol., 28: 1427 (1979).PubMedCrossRefGoogle Scholar
  38. 38.
    K. Einarsson, J.K. Gustafsson, and K. Hellstrom, Effect of clofibrate nn the maahnlism of nrnnacternna and nectradinla in rat liver microsomal fraction, Biochem. J., 136: 623 (1973).PubMedGoogle Scholar
  39. 39.
    B. Angelin, I. Bjorkhem, and K. Einarsson, Effect of clofibrate on some microsomal hydroxylations involved in the formation and metabolism of bile acids in rat liver, Biochem. J., 156: 445 (1976).PubMedGoogle Scholar
  40. 40.
    W.G. Levine, Induction and inhibition of the metabolism and biliary excretion of the azo dye carcinogen, N,N-dimethyl-4-aminoazobenzene (DAB) in the rat, Drug Metab. Dispos., 8: 212 (1980).PubMedGoogle Scholar
  41. 41.
    W.G. Levine, Effects of hypolipidemic drug nafenopin (2-methyl-2-[p(1,2,3,4-tetrahydro-1-naphthyl)phenoxy]propionic acid, TPIA, Su-13,4337) on the hepatic disposition of foreign compounds in the rat, Drug Metab. Dispos., 2: 178 (1974).PubMedGoogle Scholar
  42. 42.
    B.G. Lake, T.J.B. Gray, et al., The effect of hypolipidemic agents on peroxisomal (3-oxidation and mixed-function oxidase activities in primary cultures of rat hepatocytes. Relationship between induction of palmitoyl-CoA oxidation and lauric acid hydroxylation, Xenobiotica, 14: 269 (1984).PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1986

Authors and Affiliations

  • W. G. Levine
    • 1
  • H. Raza
    • 1
  1. 1.Department of Molecular PharmacologyAlbert Einstein College of MedicineBronxUSA

Personalised recommendations